Department of Pharmaceutical Sciences, University at Buffalo, State University of New York, Amherst, New York 14260, USA.
AAPS J. 2012 Jun;14(2):252-61. doi: 10.1208/s12248-012-9342-9. Epub 2012 Mar 6.
Our objective was to determine the pharmacokinetics, bioavailability and lymph node uptake of the monoclonal antibody bevacizumab, labeled with the near-infrared (IR) dye 800CW, after intravenous (IV) and subcutaneous (SC) administration in mice. Fluorescence imaging and enzyme-linked immunosorbent assay (ELISA) assays were developed and validated to measure the concentration of bevacizumab in plasma. The bevacizumab-IRDye conjugate remained predominantly intact in plasma and in lymph node homogenate samples over a 24-h period, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis and size exclusion chromatography. The plasma concentration vs. time plots obtained by fluorescence and ELISA measurements were similar; however, unlike ELISA, fluorescent imaging was only able to quantitate concentrations for 24 h after administration. At a low dose of 0.45 mg/kg, the plasma clearance of bevacizumab was 6.96 mL/h/kg after IV administration; this clearance is higher than that reported after higher doses. Half-lives of bevacizumab after SC and IV administration were 4.6 and 3.9 days, respectively. After SC administration, bevacizumab-IRDye800CW was present in the axillary lymph nodes that drain the SC site; lymph node uptake of bevacizumab-IRDye 800CW was negligible after IV administration. Bevacizumab exhibited complete bioavailability after SC administration. Using a compartmental pharmacokinetic model, the fraction absorbed through the lymphatics after SC administration was estimated to be about 1%. This is the first report evaluating the use of fluorescent imaging to determine the pharmacokinetics, lymphatic uptake, and bioavailability of a near-infrared dye-labeled antibody conjugate.
我们的目的是确定单克隆抗体贝伐单抗的药代动力学、生物利用度和淋巴结摄取，该抗体用近红外（IR）染料 800CW 标记，通过静脉（IV）和皮下（SC）给药后在小鼠体内。开发并验证了荧光成像和酶联免疫吸附测定（ELISA）检测法来测量贝伐单抗在血浆中的浓度。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳和尺寸排阻色谱法测定，在 24 小时内，贝伐单抗-IRDye 缀合物在血浆和淋巴结匀浆样本中主要保持完整。荧光和 ELISA 测量获得的血浆浓度-时间曲线相似；然而，与 ELISA 不同，荧光成像只能在给药后 24 小时内定量浓度。在 0.45mg/kg 的低剂量下，静脉内给药后贝伐单抗的血浆清除率为 6.96mL/h/kg；该清除率高于高剂量后报道的清除率。SC 和 IV 给药后贝伐单抗的半衰期分别为 4.6 和 3.9 天。SC 给药后，IRDye800CW 标记的贝伐单抗存在于引流 SC 部位的腋窝淋巴结中；静脉内给药后，贝伐单抗-IRDye800CW 在淋巴结中的摄取可以忽略不计。贝伐单抗经 SC 给药后表现出完全的生物利用度。使用房室药代动力学模型，估计 SC 给药后通过淋巴途径吸收的分数约为 1%。这是首次报告评估荧光成像用于确定近红外染料标记抗体缀合物的药代动力学、淋巴摄取和生物利用度。
